Masao Jinno

Measurement of endometrial tissue blood flow: a novel way to assess uterine receptivity for implantation

OBJECTIVE To assess endometrial receptivity in terms of endometrial tissue blood flow (ETBF) measured hysterofiberscopically by laser blood-flowmetry, and to examine the techniques effectiveness in an in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) program. DESIGN A prospective clinical study. SETTING(S) IVF program in a university hospital. PATIENT(S) A total of 75 infertile women with normal menstrual cycles undergoing IVF/ICSI. INTERVENTION(S) ETBF, conventional ultrasonographic, endocrinologic, and histologic parameters for receptivity and immunoreactivity for vascular endothelial growth factor (VEGF) in endometrium were assessed between days 4 and 6 of the luteal phase in a spontaneous menstrual cycle. Then all patients underwent IVF/ICSI. MAIN OUTCOME MEASURE(S) Achievement of clinical pregnancy by IVF/ICSI. RESULT(S) ETBF, VEGF expression, and the number of embryos were significantly higher in the women who became pregnant than in those who did not. By stepwise multiple logistic regression, significant predictors of pregnancy were the number of embryos and ETBF but not conventional receptivity markers. The rate of pregnancy was significantly higher in women with ETBF values of at least 29 mL/min per 100 grams of tissue than in women with lower values (42 vs. 15% in 36 and 39 women, respectively). ETBF was significantly greater in morphologically normal than abnormal uteri. In normal uteri, ETBF was greatest in the fundus. Correspondingly, in normal uteri 85% of gestational sacs were implanted in the fundus. CONCLUSION(S) ETBF is superior to conventional parameters for determining endometrial receptivity for implantation.

Complete removal of HIV-1 RNA and proviral DNA from semen by the swim-up method: Assisted reproduction technique using spermatozoa free from HIV-1

Background:Use of antiretroviral drugs has reduced the mortality rate for HIV infection and many HIV-discordant couples wish to have children. It is possible for an HIV-infected man to father children without risk of HIV transmission if HIV-free spermatozoa can be obtained from his semen. Methods:An improved swim-up method was used to collect HIV-free spermatozoa from the semen of HIV-positive males. Diluted semen was layered over a Percoll solution with a continuous density gradient of 30–98%, and then centrifuged. The bottom layer was collected by cutting the end from the tube and the sperm suspension was collected using the swim-up method. Spermatozoa were tested by nested polymerase chain reaction (PCR) for HIV-1 RNA and DNA, with a detection limit of one copy. Spermatozoa were used for assisted reproduction in 43 couples. Results:HIV-1 RNA and proviral DNA were not detected by nested-PCR assay in all 73 of the collected spermatozoa samples from 52 patients. The HIV-1-negative sperm was used for in vitro fertilization in 12 couples and for intracytoplasmic sperm injection in 31 couples. No detection of HIV-1 RNA or proviral DNA in the culture medium of the fertilized eggs was confirmed again before embryo transfer. Of the 43 female partners, 20 conceived and 27 babies were born. HIV antibodies, HIV RNA and proviral DNA were negative in all of the females and babies. Conclusions:HIV-negative spermatozoa could be obtained from semen of HIV-positive men. The method involves no risk of HIV transmission to female partners and their children.

Gonadotropin stimulates ovarian renin-angiotensin system in the rabbit

The present study was undertaken to assess the role of ovarian renin-angiotensin system (RAS) in the preovulatory cascade induced by gonadotropin exposure. In the in vitro perfused rabbit ovaries, exposure to human chorionic gonadotropin (hCG) enhanced the secretion rate of angiotensin II (Ang II) within 1 h. The secretion rate reached maximal levels at 6 h and then declined thereafter. The intrafollicular Ang II content and renin-like activity were also significantly increased at 2 and 4 h after exposure to hCG, compared with control ovaries perfused with medium alone. The level of intrafollicular Ang II after hCG exposure significantly exceeded the concentration of Ang II in an equivalent volume of plasma. The addition of 1 microM captopril to the perfusate significantly inhibited the secretion rate of Ang II stimulated by hCG; however, captopril affected neither the ovulatory efficiency nor prostaglandin production in ovaries treated with hCG. Captopril significantly inhibited the resumption of meiosis in the ovulated ova and follicular oocytes stimulated by hCG. The administration of 100 micrograms Ang II at 2-h intervals to the perfusate reversed the inhibitory effects of captopril on hCG-induced oocyte maturation. In conclusion, these data indicate that gonadotropin stimulates renin-like activity and Ang II production in the rabbit ovary. Ovarian renin-angiotensin system may play an important role in the process of oocyte maturation after exposure to gonadotropin.

Advanced glycation end-products accumulation compromises embryonic development and achievement of pregnancy by assisted reproductive technology

BACKGROUND Advanced glycation end-products (AGE) are pivotal in aging and diabetes. Aging and polycystic ovary syndrome, a diabetes-associated disease, often cause infertility. We examined how AGE accumulation affects assisted reproductive technology (ART) outcomes. METHODS In this retrospective analysis, toxic AGE (TAGE), pentosidine (Pent) and carboxymethyl lysine (CML) in blood and follicular fluid (FF) were measured in 157 ART-patients. We analyzed associations of AGE with ART outcomes and pre-ART clinical factors. RESULTS TAGE, Pent and CML in FF and TAGE in serum, showed significant negative correlations with estradiol and numbers of follicles larger than 12 mm in diameter, retrieved oocytes, fertilized oocytes and embryos. AGE, Pent in FF and TAGE in serum showed significant negative correlations with ongoing pregnancy. Areas under receiver-operating characteristic curves for AGE (0.709), Pent in FF (0.686) and TAGE in serum (0.667) were significantly larger than for the reference (0.5). Women with serum TAGE above 7.24 U/ml showed decreased oocyte numbers and ongoing pregnancy rates, even with younger age or lower Day-3 FSH. Serum TAGE correlated positively with leptin (R = 0.51), BMI, low-density lipoprotein, triglyceride, glucose, homeostasis model assessment-insulin resistance and insulin. CONCLUSIONS Serum TAGE and FF Pent accumulations correlated highly with poor follicular and embryonic development and with a lower likelihood of ongoing pregnancy. Serum TAGE predicts poor ART outcomes independent of age and Day-3 FSH.

A novel method of ovarian stimulation for in vitro fertilization: bromocriptine-rebound method

OBJECTIVE To examine whether a new method of ovarian stimulation, bromocriptine-rebound method, improves IVF outcomes compared with the conventional long protocol of GnRH agonist and hMG regimen. DESIGN A prospective clinical trial. SETTING In vitro fertilization program at a university hospital. PATIENTS Endocrine-normal ovulatory women less than 40 years of age, with normal male partners and previous failed IVF-ET using long protocol. INTERVENTIONS Patients were assigned to either bromocriptine-rebound method (group 1) or long protocol (group 2). The bromocriptine-rebound method was the same as the long protocol, except that bromocriptine was administered daily from day 4 of the preceding cycle until 7 days before hMG stimulation. MAIN OUTCOME MEASURES The number of cleaved and morphologically superior embryos, pregnancy rate per oocyte pick-up, and serum PRL concentrations during administrations of hMG. RESULTS Significantly more embryos were cleaved and had superior morphology in group 1 than group 2. Clinical and ongoing pregnancy rates per oocyte pick-up were significantly higher in group 1 (42% and 38%, respectively) than group 2 (24% and 21%, respectively). The mean PRL concentration was significantly higher in the group 1 than group 2. A significant correlation between the number of superior embryos and PRL concentrations was observed in group 1, but not in group 2. CONCLUSION The bromocriptine-rebound method enhanced embryonic development, resulting in an increased pregnancy rate compared with the long protocol.

Direct Ovarian Effect of Growth Hormone in the Rabbit

PROBLEM: This study was undertaken to assess whether growth hormone (GH) can stimulate follicle growth and ovarian steroidogenesis via putative GH receptors.

Predicting sperm retrieval rates in testicular sperm extraction for azoospermia according to endocrine profiles

Background and AimsThe present study was carried out to examine the predictive value of endocrine profiles as indicators of the sperm retrieval rate on testicular sperm extraction (TESE) in azoospermic men.MethodsPrior to TESE, the serum levels of follicle stimulating hormone (FSH), luteinizing hormone (LH), prolactin, testosterone, dihydrotestosterone (DHT), estradiol and 17 α-hydroxyprogesterone were measured and the sagittal crosssections of the testis were acquired using ultrasonography.ResultsThe sperm retrieval rates according to the cause of azoospermia were 40% for idiopathic azoospermia, and 100% for obstructive azoospermia, cryptorchidsm and ejaculatory disorder. Based on the endocrinological profiles, the sperm retrieval rates showed significant differences at 100% for FSH ≤15 mIU/mL or LH ≤2 mIU/mL, 0% for FSH > 60 mIU/mL or LH > 12 mIU/mL, and 33% for the intermediate groups (P < 0.01). Comparison of the retrieval of spermatozoa and serum DHT level for the intermediate group also showed a significant difference, with retrieval rates of 58% for DHT ≤0.5 ng/mL and 0% for DHT > 0.5 ng/mL (P < 0.01).ConclusionsThe etiology, serum FSH, LH and DHT levels are useful in predicting the sperm retrieval rates on TESE in azoospermic patients.

An improvement in the embryo quality and pregnancy rate by the pulsatile administration of human menopausal gonadotropin in patients with previous unsuccessful in vitro fertilization attempts

OBJECTIVE To examine whether the pulsatile administration of hMG improves IVF outcomes in patients with previous unsuccessful attempts using IM injections of hMG. DESIGN A prospective randomized study. SETTING In vitro fertilization program at a university hospital. PATIENTS Eighty-eight endocrine-normal ovulatory women under 40 years of age, with normal male partners and a history of unsuccessful IVF treatment by the IM administration of hMG. INTERVENTIONS Patients were assigned randomly to receive either IM (bolus group) or pulsatile administration of hMG (pulsatile group) after pituitary desensitization by a GnRH agonist. MAIN OUTCOME MEASURES The proportion of retrieved oocytes with a polar body, the number of fertilized oocytes and embryos, the proportion of morphologically superior embryos, and the rate of pregnancy per initiated cycle were compared. RESULTS The proportion of retrieved oocytes with a polar body and the number of fertilized oocytes and embryos were similar in both groups. Significantly more embryos had superior morphology in the pulsatile group (77%) than the bolus group (52%). The rates of overall and clinical pregnancy per initiated cycle were significantly higher in the pulsatile group (39% and 30%, respectively, n = 44) than in the bolus group (18% and 11%, respectively, n = 44). CONCLUSION In women with failed IVF attempts using IM administration of hMG, the pulsatile administration of hMG produces superior embryos and, hence, a higher pregnancy rate.

Administration of human chorionic gonadotropin for in vitro fertilization-embryo transfer based on the serum luteinizing hormone (LH) concentration: the importance of synchronization with endogenous LH rises

OBJECTIVE To examine whether synchronized administration of hCG at the onset of the endogenous LH rise promotes successful IVF. DESIGN A prospective randomized study. SETTING In vitro fertilization program at a university hospital. PATIENTS A total of 208 IVF cycles in 148 patients. INTERVENTIONS Serum LH concentrations were measured daily and hMG was administered daily. Independent of follicle size and E2 concentration, hCG was administered as soon as the LH concentration exceeded the J level, defined as the minimum value + (the day 3 value-the minimum value) x 1/3(J group). Alternatively, hCG was administered when the serum LH concentration turned to increase but was still less than the J level, or 1 day after the serum LH concentration exceeded the J level (non-J group). RESULTS The rates of total and ongoing pregnancy per cycle were significantly higher in the J group (35.6% and 26.0%, respectively, n = 104) than in the non-J group (21.2% and 12.5%, respectively, n = 104). Pregnancies in the J group were achieved over a wide range of dominant follicle diameters (13 to 25 mm), E2 levels (198 to 1,700 pg/mL; conversion factor to SI units, 3.671), and E2 level per follicle > or = 12 mm (24 to 225 pg/mL per follicle) recorded on the day of hCG administration. CONCLUSION Synchronized administration of hCG in accordance with endogenous LH rises produces a high rate of pregnancy in IVF.